Flowcytometric Based Analysis of Sox4 Gene Expression-An Early Transcription Factor Influenced by 5-Azacytidine and Compare with EpCAM in Circulating Tumor Cells Isolated from Breast Cancer Patients

Circulating tumor cells (CTCs) are the frontier area of cancer medicine for early diagnosis during the management of breast cancer (BC) patients.Flowcytometry is a highly sensitive and powerful technique used for quantitative analysis of epithelialmesenchymal transition (EMT) expression of Sox4 and EpCAM using monoclonal antibodies in CTCs population, conjugated by PE (phycoerythrin) that detects more than 98.90 % score after gating.Sox4, is an early transcription factor regulating various developmental process including EMT during tumor progression.The mean values of Sox4 showing variation between -ve mAbSox4 (7427.8)and +ve mAbSox4 (6482.3),while, highest frequency (40.00%) were observed between 51-60 years age-group and O.R (4.0) with confidence interval (C.I) at 95% varying between 1.074 -14.896 showing significant (p<0.002)differences.EpCAM mean values varying between -ve mAbEpCAM (22050.8)and +ve mAbEpCAM (21251.5)and highest frequency (40.00 %) were observed between 51-61 years age-groups.Significant (p<0.003)differences with O.R (2.0) and C.I at 95% that varying between 1.074 -14.896, suggesting confirm signaling of Sox4 act as transition inducer for CTCs in BC patients.Sensitivity was further evaluated after in-vitro exposure (1.0 ugm/ml) to peripheral blood lymphocytes (PBL) with 5-azacytidine (5AzaC) and compare with controls, that showing increased mean CTCs population in 5AzaC +ve mAbSox4 (302397) and -ve mAbSox4 (251443).Statistical analysis showing observed values of O.R (1.11) and C.I at 95% varies between 0.225 -5.46, showing lack of significance (p<0.089)differences but data of the EpCAM showing decreasing trend of cell population with significant (p<0.001)differences, suggesting, EpCAM marker seems to be more sensitive EMT marker during tumorogenesis in BC.